AbstractA selective and sensitive novel methods were developed and validated for identification and trace level quantification (0.5 ppm) of genotoxic impurities Methyl 3‐amino‐4‐methylbenzoate, 3‐amino‐4‐methylbenzoic acid, and 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline in Nilotinib dihydrochloride active pharmaceutical ingredient by using liquid chromatography‐tandem mass spectrometry. The developed and validated methods were more accurate and capable to confirm the m/z values of parent and fragment ions through mass spectrometry and tandem mass spectrometry for further fragmentation. In addition, the developed methods were validated through current regulatory guidelines with a lower detection level of 0.15 ppm and a quantification level of 0.5 ppm for all the three genotoxic impurities. The correlation coefficient was observed as 0.9997 for Methyl 3‐amino‐4‐methylbenzoate genotoxic impurity‐I, 0.9998 for 3‐amino‐4‐methylbenzoic acid, and > 0.9999 for 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline. The recovery percentage of all the three genotoxic impurities was found to be between 93% to 105%.